Product Name: EGFR-pY869
Product Number: AB-PK602
Size: 25 µg      Price:89.00
Target Full Name: Epidermal growth factor receptor-tyrosine kinase

Target Alias: EGFR; Epidermal growth factor receptor; ErbB-1; ErbB, mENA; HER1; Receptor tyrosine-protein kinase ErbB-1; V-erb-b oncogene homologue; PIG61; CCDS5514.1; ENSG00000146648

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK602

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y869

Protein UniProt: P00533

Protein SigNET: P00533

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human EGFR (ErbB1) sequence peptide Cat. No.: PE-04AFS95

Antibody Immunogen Sequence: EKE(pY)HAEG(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues E866 to G873; In the protein kinase catalytic domain.

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Antibody Concentration: 1 mg/ml

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 2 µg/ml for immunoblotting

Antibody Potency: Weak immunoreactivity of a target-sized protein by Western blotting in 1 minute EGF-treated A431 cells (SDS-PAGE buffer extracted). Very strong immunoreactivity with immunogen peptide on dot blots.

Antibody Species Reactivity: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 160-180 kDa.

Antibody Specificity: Very high

Antibody Cross Reactivity: No significant cross-reactivities detected in A431, A549 and HepG2 cells.

Related Product 1: EGFR-pY869 blocking peptide

Related Product 2: EGFR-pY998 phosphosite-specific antibody (Cat. No.: AB-PK603)

Related Product 3: EGFR-pY1069 phosphosite-specific antibody (Cat. No.: AB-PK599)

Related Product 4: EGFR-pY1110 phosphosite-specific antibody (Cat. No.: AB-PK600)

Scientific Background: EGFR (ERBB1, HER1) is a protein-tyrosine kinase of the TK group and EGFR family. EGFR regulates cell proliferation, division, motility, survival, and has other roles in tissue development. It is activated by binding epidermal growth factor (EGF) and transforming growth factor-alpha (TGFa), which induce dimerization and autophosphorylation. Phosphorylation of Y270 stimulates interaction with EGFR. Phosphorylation of Y869 increases phosphotransferase activity and induces interaction with COX2. Phosphorylation of Y1016 increases phosphotransferase activity and induces interaction with GrbB2, PLCg1, PTPN11 (SHP2), RasGAP, and Vav2. Phosphorylation of Y1069 increases phosphotransferase activity and induces interaction with Cbl. Phosphorylation of Y1092 induces interaction with PLCg1, PTPN6 (SHP1) and Ras-GAP. Phosphorylation of Y1110 induces interaction with Ras-GAP. Phosphorylation of Y1172 increases phosphotransferase activity and induces interaction with Dok1, EGFR, RasGAP, PTPN11 (SHP2), and Vav2. Phosphorylation of Y1197 increases phosphotransferase activity and induces interaction with Cbl, EGR, PLCg1, RasGAP, SH3KBP1, Shc1, and PTPN6 (SHP1). EGFR is inhibited by phosphorylation at T678,S695 and S1026. Phosphorylation of T693, S695, S1070, S1071, and S1190 contributes to receptor internalization. EGFR is a known oncoprotein (OP). Cancer-related mutations in human tumours point to a gain of function of the protein kinase. Constitutive activation of EGFR kinase activity has been seen with the mutations of V689M, E1005R+D1006K. The active form of the protein kinase normally acts to promote tumour cell proliferation.
Figure 1. Dot blotting EGFR-pY869 antibody with recombinant purified proteins.

Figure 2. Epitope mapping of EGFR-pY869 antibody with similar phosphopeptides on dot blots.

Figure 3. Identification of phosphosites related to EGFR-pY869.